臭椿枝叶化学成分抗烟草花叶病毒活性研究

吴江梅; 左安建; 邹正彪; 刘洋; 张威; 符滔; 闫晓慧; 胡世俊

doi:10.19303/j.issn.1008-0384.2023.04.009

臭椿枝叶化学成分抗烟草花叶病毒活性研究

doi: 10.19303/j.issn.1008-0384.2023.04.009

吴江梅^1,,
左安建¹,
邹正彪¹,
刘洋¹,
张威¹,
符滔¹,
闫晓慧^1, ,,
胡世俊^2, ,

1.
西南林业大学生物多样性保护学院/云南省森林灾害预警与控制重点实验室，云南　昆明　650224
2.
西南林业大学林学院，云南　昆明　650224

基金项目: 国家自然科学基金项目（32160378）；云南省应用基础研究重点项目（2019FA011）；云南省高层次人才项目（YNWR-QNBJ-2018-300）

详细信息

作者简介:
吴江梅（1997−），女，硕士研究生，研究方向：农业有害生物综合防控（E-mail：1239350410@qq.com）

通讯作者:
闫晓慧（1980−），女，博士，教授，研究方向：植物活性成分（E-mail：luckyyxh@163.com）

胡世俊（1977−），男，博士，副教授，研究方向：入侵植物（E-mail：shijunhu@126.com）

中图分类号: S482.1
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出版历程
- 收稿日期: 2022-10-17
- 修回日期: 2023-02-25
- 网络出版日期: 2023-05-09
- 刊出日期: 2023-04-28

Anti-TMV Activity of Chemicals in Leaves and Branches of Ailanthus altissima

1.
Key Laboratory of Forest Disaster Warning and Control in Yunnan Province/College of Biodiversity Conservation, Southwest Forestry University, Kunming, Yunnan　650233, China
2.
College of Forestry，Southwest Forestry University, Kunming, Yunnan　650233, China

摘要

摘要: 目的对臭椿（Ailanthus altissima）化学成分及其抗烟草花叶病毒（TMV）活性进行研究，为开发新型植物病毒抑制剂提供理论依据。方法综合运用硅胶、凝胶、MCI等多种柱层析方法对臭椿枝叶正丁醇提取物化学成分进行分离，利用NMR、MS鉴定其结构；以TMV为供试病毒，采用半叶枯斑法从保护活性、治疗活性、钝化活性等3个方面评估化合物的生物活性。结果从臭椿枝叶正丁醇提取物中分离鉴定了17个化合物，根据其理化性质以及波谱数据分别鉴定为：山奈酚（Kaempferol） (1)、(2S)-3-O-Octadeca-9Z,12Z,15Z-trienoylgycery-O-β-D-galactopyranoside (2)、正二十六烷（Hexacosane） (3)、6,9,12-Octadecatrienoic acid (4)、Eichlerianic acid (5)、 Colocasinol A (6)、咖啡酸二十烷酯（Caffeic acid eicosanyl ester） (7)、Acernikol (8)、(-)-Sakuyayesinol (9)、(14S,17S,20S,24R)-20,24,25-trihydroxy-14,17-cylomalabarican-3-one (10)、Pinnata E(11)、morin-3-O-α-rhamnopyranoside(12)、Trans-syringin (13)、姜糖脂A（Gingerglycolipid A） (14)、姜糖脂B（gingerglycolipid B）(15)、benzyl 2-O-β-apiofuranosyl-(1→2)-β-D-glucopyranosyl-2,6-dihydroxy-benzoate (16)、picrorhizoside C (17)。化合物2、14、15为首次从该植物中分离得到。在质量浓度为50 μg·mL^-1时，化合物6、8、13、14对TMV的钝化活性较为显著，抑制率均在50%以上，与阳性对照药剂宁南霉素无显著差异，分别为木脂素类化合物、木脂素类化合物、苯丙素类化合物和半乳糖脂类化合物。结论从臭椿枝叶中分离得到的17个化合物，均对TMV具有抑制作用，部分半乳糖脂类化合物、木脂素类化合物、苯丙素类化合物对TMV有显著的钝化作用，研究结果丰富了臭椿抗TMV活性物质的范畴，也为今后开发新型植物病毒抑制剂提供了科学依据。
- 臭椿 /
- 化学成分 /
- 分离鉴定 /
- 半叶枯斑法 /
- 抗TMV活性
Abstract: Objective Chemical composition and anti-tobacco mosaic virus (TMV) activity of Ailanthus altissima were studied to pave the way for developing an effective viral inhibitor. Methods Substrates in the n-butanol extract of leaves and branches of A.altissima plants were separated using silica gel, Sephadex LH-20, and MCI column chromatography.Chemical structures of the substrates were determined based on NMR and MS data, and anti-TMV activity of the compounds examined using the half-leaf method. Results The 17 compounds isolated from the n-butanol extract were identified to be: (1) kaempferol, (2) (2S)-3-o-octadeca-9Z, 12Z, 15Z-trienoylgycery-O-β-D-galactopyranoside, (3) hexacosane, (4) 6,9,12-octadecatrienoic acid, (5) eichlerianic acid, (6) colocasinol A, (7) caffeic acid eicosanyl ester, (8) acernikol, (9) (-)-Sakuyayesinol (10), (14S,17S, 20S, 24R)-20,24,25-trihydroxy-14,17-cylomalabarican-3-one, (11) 4-(3-butoxy-1-hydroxy-2-methoxypropyl) benzene-1, 2-diol, (12) pinnata, (13) trans-syringin, (14) gingerglycolipid A, (15) gingerglycolipid B, (16) benzyl 2-o-β-apiofuranosyl-(1→2)-β-D-glucopyranosyl-2,6-dihydroxy-benzoate, and (17) picrorhizoside C.Among them, Compounds 2, 14, and 15 were isolated from the plant for the first time.And the lignans, phenylpropanoids, or galactose lipids, such as Compounds 6, 8, 13, and 14 at concentration of 50 μg·mL⁻¹ displayed a TMV inactivation rate greater than 50%, which was similar to that of the positive control, ningnamycin.For Compounds 6, 13, and 14, the rates were even higher than that of ningnamycin. Conclusion All 17 substances isolated from the leaves and branches of A.altissima exhibited varying degrees of inhibitory effect on TMV.The efficacy was more significant as shown by certain Lignans, phenylpropanoids, and galactose lipids.
- Ailanthus altissima /
- chemical composition /
- isolation and identification /
- half-leaf method /
- anti-TMV activity

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图 1 化合物1-17的分离流程

Figure 1. Flow diagram on separating chemicals in n-butanol extract of A.altissima leaves and branches

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图 3 化合物2、7和11对TMV侵染的治疗效应

a：化合物2 的治疗作用；b：化合物7的治疗作用；c：化合物11的治疗作用。

Figure 3. Curative effects of Compounds 2, 7, and 11 against TMV infection

a: Curative effect of Compound 2; b: Curative effect of Compound 7; c: Curative effect of Compound 11;

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图 4 化合物6、13和14对TMV侵染的钝化作用

a化合物6 的钝化作用；b化合物13 的钝化作用；c化合物14的钝化作用

Figure 4. TMV-inactivation effects of Compounds 6, 13, and 14

a: TMV-inactivation effect of Compound 6; b: inactivation effect of Compound 13; c: inactivation effect of compound 14.

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表 1 化合物1～17对TMV的保护作用

Table 1. Anti-TMV effects of 17 compounds

化合物 Compound	处理平均枯斑数 Local lesions of treatment	对照平均枯斑数 Local lesions of control	P值 P value	抑制率 Inhibition rate/%
1	18.33±5.51	24.00±7.55	0.35	23.42±1.63 g
2	16.33±8.02	24.67±12.01	0.37	34.25±4.71 cd
3	12.33±4.04	14.33±5.03	0.62	13.41±2.34 h
4	13.33±4.04	21.00±7.21	0.18	35.65±4.35 bc
5	17.67±6.51	23.00±7.55	0.41	23.89±3.47 g
6	14.33±7.09	22.00±12.12	0.40	33.14±5.19 cd
7	19.67±4.04	29.00±5.57	0.07	32.30±0.96 de
8	22.00±7.00	29.33±9.87	0.35	24.56±2.16 fg
9	12.67±7.37	17.67±10.02	0.52	28.72±1.21 ef
10	14.67±6.51	24.33±11.06	0.26	39.77±4.36 b
11	15.67±7.02	25.00±11.53	0.30	37.02±1.14 bc
12	27.33±14.74	31.00±15.87	0.78	12.66±2.85 h
13	20.33±5.03	31.00±9.17	0.16	33.80±5.78 cd
14	12.33±4.16	19.00±4.58	0.14	36.02±5.96 bc
15	11.33±5.69	15.67±6.66	0.44	29.69±8.90 de
16	11.00±3.00	15.00±4.00	0.24	26.75±0.48 ef
17	13.00±6.00	18.33±9.50	0.46	27.38±4.97 ef
宁南霉素（Ningnanmycin）	3.67±1.73	8.33±3.79	0.02	55.19±1.30 a
数据为3次重复取平均值；化合物质量浓度为50 μg·mL⁻¹；不同字母表示在5%水平显著，下同。 Data are averages of triplicate; mass concentration at 50 μg·mL⁻¹; data with different letters indicate significance at 5% level.Same for below.

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表 2 化合物1～17对TMV的治疗作用

Table 2. Anti-TMV effects of 17 compounds on infected tissue

化合物 Compound	处理平均枯斑数 Local lesions of treatment	对照平均枯斑数 Local lesions of control	抑制率 Inhibition rate/%
1	17.33±4.73	22.67±5.86	23.74±1.41 f
2	16.67±8.33	31.00±15.13	46.45±0.81 b
3	16.00±6.83	22.25±10.40	27.06±3.33 f
4	13.33±6.81	17.67±8.33	25.39±2.76 f
5	13.33±4.04	17.67±5.13	24.68±1.82 f
6	17.67±6.51	24.00±8.54	26.56±1.50 f
7	23.33±6.35	34.33±10.02	31.70±3.56 ef
8	19.00±3.00	26.67±5.03	28.48±2.40 f
9	14.00±6.56	21.33±10.07	34.24±0.84 de
10	26.33±4.51	41.67±8.62	36.50±2.35 d
11	18.67±3.79	33.33±7.57	43.81±1.26 c
12	16.50±5.26	31.50±10.41	47.11±4.21 b
13	23.00±4.00	28.67±5.69	19.50±4.18 g
14	14.33±8.02	19.00±11.00	24.24±2.88 f
15	26.00±7.55	36.33±10.50	28.38±1.92 f
16	12.67± 5.51	15.33±6.66	17.41±0.76 g
17	10.33± 4.16	14.00±6.25	25.26±3.18 f
宁南霉素（ningnanmycin）	16.00± 2.00	34.33±6.51	53.01±3.05 a

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表 3 化合物1-17对TMV的钝化作用

Table 3. TMV inactivation effects of 17 compounds

化合物 Compound	处理平均枯斑数 Local lesions of treatment	对照平均枯斑数 Local lesions of control	P值 P value	抑制率 Inhibition rate/%
1	12.33±3.51	27.67±6.43	0.02	55.72±4.61 b
2	21.33±3.79	29.00±6.08	0.13	26.04±2.9 ef
3	14.67±3.51	22.00±7.00	0.18	32.14±5.64 de
4	9.33±4.51	15.00±7.55	0.32	37.45±1.71 cd
5	13.67±7.51	20.67±12.01	0.44	33.23±3.18 de
6	10.33±3.51	28.33±7.51	0.02	64.02±2.79 a
7	22.00±6.25	40.33±9.29	0.04	45.86±3.64 c
8	22.33±5.69	53.00±13.53	0.02	57.87±2.01 ab
9	19.00±2.00	30.00±4.36	0.02	36.33±4.31 cd
10	24.67±8.33	42.00±10.82	0.09	42.03±4.42 c
11	14.67±3.21	26.33±5.51	0.03	44.39±0.92 c
12	26.33±10.02	40.00±16.52	0.28	33.65±3.05 de
13	9.00±1.00	26.00±3.61	0.001	65.18±3.79 a
14	16.33±3.21	45.33±6.81	0.002	63.40±9.21 ab
15	25.67±1.53	31.67±3.06	0.03	18.74±3.61 f
16	27.00±8.19	43.67±15.04	0.16	37.55±3.32 cd
17	20.00±3.61	30.67±5.51	0.04	34.77±2.47 cd
宁南霉素（ningnanmycin）	5.67±1.53	15.00±1.53	0.07	61.72±1.98 ab

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